Académique Documents
Professionnel Documents
Culture Documents
Irving Center for Clinical Research, Columbia University College of Physicians and Surgeons, 630 W 168th St., New York 10032, New York, USA
Department of Medicine, Columbia University College of Physicians and Surgeons, 630 W 168th St., New York 10032, New York, USA
New York Hospital-Cornell Medical Center, White Plains, New York 10605, New York, USA
Department of Pathology, Columbia University College of Physicians and Surgeons, 630 W 168th St., New York 10032, New York, USA
(Requests for offprints should be addressed to J F OConnor, Department of Pathology and Irving Center for Clinical Research, Columbia University College of
Physicians and Surgeons, 630 W 168th St., PH10305, New York 10032, New York, USA)
Abstract
Human chorionic gonadotropin (hCG) exhibits molecular
heterogeneity in both its protein and carbohydrate
moieties. This communication describes changes in hCG
isoforms detected directly in clinical samples. These isoforms, quantified in blood or urine specimens, show a
progression of change throughout normal pregnancy. Early
pregnancy produces a type of hCG that resembles, in
terms of immunoreactivity, a major form of hCG excreted
in choriocarcinoma. The isoforms predominate for the first
56 weeks of gestation and then diminish, being replaced
with the hCG isoforms which predominate throughout
the remainder of pregnancy. The alteration in hCG
isoform content occurs in both blood and urine. The
progression of isoforms is best delineated by calculating the
Introduction
Human chorionic gonadotropin (hCG) is produced by the
placental trophoblast of normal pregnancy, and in gestational trophoblastic disease (hydatidiform mole and choriocarcinoma) (Hussa 1987). It is also produced in much smaller
quantities by the pituitary (Birken et al. 1996) in both preand postmenopausal women and in men (Odell et al. 1990)
and also in many non-gestational malignant tumors (Hussa
1987).
Structural variations in both the protein and carbohydrate portions of the hCG molecule have been well
established. They may occur as a concomitant of its origin
(Wide & Hobson 1987, Birken et al. 1996), or metabolic
transformation (Nisula et al. 1989). The most prominent
differences documented to date are between the hCG
produced by choriocarcinoma and that of mid-first trimester normal pregnancy, with choriocarcinoma-derived
hCG having a generally higher content of more highly
branched oligosaccharides (Cole 1987, Elliott et al. 1997).
Even in normal pregnancy, multiple isoforms of hCG
exist and their relative amounts vary as pregnancy
100
G KOVALEVSKAYA
and others
IRMAs
The methodology used in the construction and validation
of the B109B108* assay has been fully described elsewhere (OConnor et al. 1988); the first antibody is the
capture antibody, the second, with an asterisk, is a
radioiodinated detection antibody. The B152B207* assay
has also been characterized (OConnor et al. 1998). Both
assays were performed with a slight modification of the
published procedure: the capture antibody was adsorbed
onto the wells of microtiter plates (Immulon IV,
Dynatech, Chantilly, VA, USA) by incubating a 5 g/ml
solution (B109B108* assay) or 25 g/ml solution (B152
Journal of Endocrinology (1999) 161, 99106
Patient samples
Urine samples from in vitro fertilization (IVF) patients were
a gift from Dr L Cole. They included spontaneous abortion
(n=14, range of gestational age 1841 weeks from
embryo transfer (ET)), ectopic pregnancies (n=7, gestational age 2340 weeks) and normal pregnancy controls
(n=65, encompassing the range 0654 weeks from ET).
Some of the normal pregnancy urine samples throughout
the pregnancies were also obtained from Dr Cole. Others
were obtained from the clinical practice of collaborating
physicians at Columbia Presbyterian Medical Center
G KOVALEVSKAYA
and others
isoforms ranged from 62 to 13, indicating a predominance of the B152 isoform(s) in early pregnancy. During
the 1012 week period, the ratio ranged from 10 to 02,
indicating that an inversion in hCG isoform content is
occurring as pregnancy progresses. This decline in the
ratio continues, ranging from 054 to 008 in the 1518
week period and reaching an inflection point at 29 weeks.
Journal of Endocrinology (1999) 161, 99106
101
102
G KOVALEVSKAYA
and others
Measure
Serum, ratio B152/B109
Serum, log(ratio B152/B109)
Urine, ratio B152/B109
Urine, log(ratio B152/B109)
t-test=(df)
t(11) =665
t(23) =2161
t(11) =464
t(157)=1685
00001
00000
00007
00001
G KOVALEVSKAYA
and others
Table 2 Analysis of the B152/B109 ratio in serum vs urine in the first and third trimesters
of pregnancy
Measure
Paired t (df)
Ratio B152/B109
Log(ratio B152/B109)
t(11)=325
t(11)=625
00077
00001
Ratio B152/B109
Log(ratio B152/B109)
t(10)=547
t(10)=714
00003
00001
103
104
G KOVALEVSKAYA
and others
Table 3 IVF patients: analysis of covariance of hCG isoforms among normal pregnancy,
ectopic pregnancy and spontaneous abortion as a function of gestational age
Adjusted r2c
Pairwise differenced
Diagnosis
Fe
Outcome
Log(ratio B152/B109)a
051
089
056
2133
00001
Log(B152)/log(B109)b
045
434
0016
Log(B152B207*)b
050
2694
00001
Log(B109B108*)
041
None
Normal pregnancy vs
abortion and ectopic
Normal pregnancy vs
abortion
Normal pregnancy vs
abortion and ectopic
G KOVALEVSKAYA
and others
105
106
G KOVALEVSKAYA
and others
Ulloa-Aguirre A, Mendez JP, Cravioto A, Grotjan E, DamianMatsumura P & Espinoza R 1990 Studies on the microheterogeneity
of chorionic gonadotrophin secreted by the human cytotrophoblast in
culture. Human Reproduction 5 661669.
Vaitukaitis JL, Braunstein GD & Ross GT 1972 A radioimmunoassay
which specifically measures human chorionic gonadotropin in the
presence of human luteinizing hormone. American Journal of
Obstetrics and Gynecology 113 751758.
Wide L & Hobson B 1987 Some qualitative differences of hCG in
serum from early and late pregnancies and trophoblastic diseases.
Acta Endocrinologica 116 465472.
Wide L, Lee JY & Rasmussen C 1994 A change in the isoforms of
human chorionic gonadotropin occurs around the 13th week of
gestation. Journal of Clinical Endocrinology and Metabolism 78
14191423.